An indirect competitive enzyme-linked immunosorbent assay toward the standardization of Pueraria candollei based on its unique isoflavonoid, kwakhurin

Gorawit Yusakul, Rui Togita, Kei Minami, Kaskamol Chanpokapaiboon, Thaweesak Juengwatanatrakul, Waraporn Putalun, Hiroyuki Tanaka, Seiichi Sakamoto, Satoshi Morimoto

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Kwakhurin (Kwa) is a plant secondary metabolite solely present in Pueraria candollei var. mirifica (P. candollei), which has long been used as a Thai traditional herb for estrogen replacement therapy. Recently, health hazards have arisen in Japan regarding P. candollei-derived products containing potent estrogenic compounds. Therefore, the development of standardization methods for P. candollei materials is an urgent problem requiring resolution. The enzyme-linked immunosorbent assay (ELISA) is an effective analytical technique because it enables the development of sensitive and specific assays of the target compound through antigen−antibody reaction. Here, we produced a monoclonal antibody against Kwa (MAb 11F) by immunizing Kwa-bovine serum albumin (BSA) conjugates prepared using an N,N′-carbonyldiimidazole (CDI) mediated method. Stability and cross-reactivity tests of MAb 11F revealed that the MAb 11F is stable for at least 4 months at 4 °C and is highly specific to Kwa. The detectable concentration range of an indirect competitive ELISA (icELISA) using MAb 11F exhibited values of 1.53–48.8 ng/mL with the limit of detection (LOD) of 1.13 ng/mL. Validation analyses revealed that the developed icELISA is precise, accurate, and reliable enough to be applied to P. candollei-derived samples and products for their standardization.

Original languageEnglish
Pages (from-to)23-28
Number of pages6
JournalFitoterapia
Volume133
DOIs
Publication statusPublished - Mar 2019

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Drug Discovery

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